Method of Treating Cancer

ABSTRACT

This invention is directed to the treatment of cancer, particularly castration-resistant prostate cancer and osteolytic bone metastases, with a dual inhibitor of MET and VEGF.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a United States National Phase filing ofPCT/US2013/039284, filed May 2, 2013, which claims the benefit ofpriority of U.S. Provisional Application No. 61/641,837, filed May 2,2012, all of which are incorporated herein by reference.

FIELD OF THE INVENTION

This invention is directed to the treatment of cancer, particularlycastration-resistant prostate cancer and osteolytic bone metastases,with a dual inhibitor of MET and VEGF.

BACKGROUND OF THE INVENTION

Castration-Resistant Prostate Cancer (CRPC) is a leading cause ofcancer-related death in men. Despite progress in systemic therapy forCRPC, improvements in survival are modest, and virtually all patientssuccumb to this disease within about 2 years. The primary cause ofmorbidity and mortality in CRPC is metastasis to the bone, which occursin about 90% of cases.

Metastasis to the bone is a complex process that involves interactionsbetween cancer cells and components of the bone microenvironmentincluding osteoblasts, osteoclasts, and endothelial cells. Bonemetastases cause local disruption of normal bone remodeling, and lesionsgenerally show a propensity for either osteoblastic (bone-forming) orosteolytic (bone-resorbing) activity. Although most CRPC patients withbone metastases display features of both types of lesions, prostatecancer bone metastases are often osteoblastic, with abnormal depositionof unstructured bone accompanied by increased skeletal fractures, spinalcord compression, and severe bone pain.

The receptor tyrosine kinase MET plays important roles in cell motility,proliferation, and survival, and it has been shown to be a key factor intumor angiogenesis, invasiveness, and metastasis. Prominent expressionof MET has been observed in primary and metastatic prostate carcinomas,with evidence for higher levels of expression in bone metastasescompared to lymph node metastases or primary tumors.

Vascular endothelial growth factor (VEGF) and its receptors onendothelial cells are widely accepted as key mediators in the process oftumor angiogenesis. In prostate cancer, elevated VEGF in either plasmaor urine is associated with shorter overall survival. VEGF may also playa role in activating the MET pathway in tumor cells by binding toneuropilin-1, which is frequently unregulated in prostate cancer andappears to activate MET in a co-receptor complex. Agents targeting theVEGF signaling pathway have demonstrated some activity in patients withCRPC.

MET signalling can influence osteoblast and osteoclast function. Strongimmunohistochemical staining of MET has been observed in osteoblasts indeveloping bone, while both HGF and MET are expressed by osteoblasts andosteoclasts in vitro and regulate cellular responses such asproliferation, migration and differentiation. Secretion of HGF byosteoblasts has been proposed as a key factor in osteoblast/osteoclastcoupling [58] and is thought to promote the development of bonemetastases by tumour cells that express MET.

Like MET, the VEGF signalling pathway is strongly implicated in boneformation and remodelling. Both osteoblasts and osteoclasts express VEGFand VEGF receptors, which appear to be involved in autocrine and/orparacrine feedback mechanisms regulating cell proliferation, migration,differentiation and survival. Experiments using genetically modifiedmice have shown that angiogenesis and VEGF signalling in osteoblasts areboth important in bone development and repair.

Thus, a need remains for methods of treating prostate cancer, includingCRPC and the associated osteolytic bone metastases.

SUMMARY OF THE INVENTION

These and other needs are met by the present invention which is directedto a method for treating bone cancer associated with prostate cancer.The method comprises administering a therapeutically effective amount ofa compound that modulates both MET and VEGF to a patient in need of suchtreatment. In one embodiment, the bone cancer is osteolytic bonemetastases. In a further embodiment, the prostate cancer is CRPC. In afurther embodiment, the bone cancer is osteolytic bone metastasesassociated with CRPC.

In one aspect, the present invention is directed to a method fortreating osteolytic bone metastases, CRPC, or osteolytic bone metastasesassociated with CRPC, comprising administering a therapeuticallyeffective amount of a compound that dually modulates MET and VEGF to apatient in need of such treatment.

In one embodiment of this and other aspects, the dual acting MET/VEGFinhibitor is a compound of Formula I

or a pharmaceutically acceptable salt thereof, wherein:

R¹ is halo;

R² is halo;

R³ is (C₁-C₆)alkyl;

R⁴ is (C₁-C₆)alkyl; and

Q is CH or N.

In another embodiment, the compound of Formula I is a compound ofFormula Ia

or a pharmaceutically acceptable salt thereof, wherein:

R¹ is halo;

R² is halo; and

Q is CH or N.

In another embodiment, the compound of Formula I is compound 1:

or a pharmaceutically acceptable salt thereof. Compound 1 is known asN-(4-{[6,7-bis(methyloxy)quinolin-4-yl]oxy}phenyl)-N′-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide.

In another embodiment, the compound of Formula I, Ia, or Compound 1 isadministered as a pharmaceutical composition comprising apharmaceutically acceptable additive, diluent, or excipient.

In another aspect, the invention provides a method for treatingosteolytic bone metastases associated with CRPC, comprisingadministering a therapeutically effective amount of a pharmaceuticalcomposition comprising Compound of Formula I or the malate salt ofCompound of Formula I or another pharmaceutically acceptable salt ofCompound of Formula I, to a patient in need of such treatment. In aspecific embodiment, the Compound of Formula I is Compound 1.

In another aspect, the invention provides a method for reducing orstabilizing metastatic bone lesions associated with CRPC, comprisingadministering a therapeutically effective amount of a pharmaceuticalcomposition comprising Compound of Formula I, Ia or the malate salt ofCompound of Formula I or another pharmaceutically acceptable salt ofCompound of Formula I, to a patient in need of such treatment. In aspecific embodiment, the Compound of Formula I is Compound 1.

In another aspect, the invention provides a method for reducing bonepain due to metastatic bone Lesions associated with CRPC, comprisingadministering a therapeutically effective amount of a pharmaceuticalcomposition comprising Compound of Formula I or the malate salt ofCompound of Formula I or another pharmaceutically acceptable salt ofCompound of Formula I, to a patient in need of such treatment. In aspecific embodiment, the Compound of Formula I is Compound 1.

In another aspect, the invention provides a method for treating orminimizing bone pain due to metastatic bone lesions associated withCRPC, comprising administering a therapeutically effective amount of apharmaceutical composition comprising Compound of Formula I or themalate salt of Compound of Formula I or another pharmaceuticallyacceptable salt of Compound of Formula I, to a patient in need of suchtreatment. In a specific embodiment, the Compound of Formula I isCompound 1.

In another aspect, the invention provides a method for strengtheningbones in patients with metastatic bone lesions associated with CRPC,comprising administering a therapeutically effective amount of apharmaceutical composition comprising Compound of Formula I or themalate salt of Compound of Formula I or another pharmaceuticallyacceptable salt of Compound of Formula I, to a patient in need of suchtreatment. In a specific embodiment, the Compound of Formula I isCompound 1. Bone strengthening can occur when the disruption in normalbone remodeling due to bone metastases is minimized, for instance byadministering a Compound of Formula I as provided herein.

In another aspect, the invention provides a method for preventingosteolytic bone metastases associated with CRPC, comprisingadministering a therapeutically effective amount of a pharmaceuticalcomposition comprising Compound of Formula I or the malate salt ofCompound of Formula I or another pharmaceutically acceptable salt ofCompound of Formula I, to a patient in need of such treatment. In aspecific embodiment, the Compound of Formula I is Compound 1.

In another aspect, the invention provides a method for preventingosteolytic bone metastases in patients with prostate cancer who arecastration resistant but have not yet advanced to metastatic disease,comprising administering a therapeutically effective amount of apharmaceutical composition comprising Compound of Formula I or themalate salt of Compound of Formula I or another pharmaceuticallyacceptable salt of Compound of Formula I, to a patient in need of suchtreatment. In a specific embodiment, the Compound of Formula I isCompound 1.

In another aspect, the invention provides a method for extending theoverall survival in patients with CRPC with osteolytic bone metastases,comprising administering a therapeutically effective amount of apharmaceutical composition comprising Compound of Formula I or themalate salt of Compound of Formula I or another pharmaceuticallyacceptable salt of Compound of Formula I, to a patient in need of suchtreatment.

In these and other aspects, the ability of the compound of Formula I totreat, ameliorate, or reduce the severity of bone metastases can bedetermined both qualitatively and quantitatively using variousphysiological markers, such as circulating tumor cell (CTC),Cross-linked C-terminal telopeptides of type-1 collagen (CTx), andCross-linked N-terminal telopeptides of type-1 collagen (NTx) counts andimaging technologies. The imaging technologies include positron emissiontomography (PET) or computerized tomography (CT) and magnetic resonanceimaging. By using these imaging techniques, it is possible to monitorand quantify the reduction in tumor size and the reduction in the numberand size of bone lesions in response to treatment with the compound ofFormula I.

In these and other aspects, shrinkage of soft tissue and viscerallesions has been observed result when the compound of Formula I isadministered to patients with CRPC. Moreover, administration of thecompound of Formula I leads to increases in hemoglobin concentration inpatients CRPC patients with anemia.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A-C show the bone scan (FIG. 1A), bone scan response (FIG. 1B),and CT scan data (FIG. 1C) for Patient 1.

FIGS. 2A-C show the bone scan (FIG. 2A), bone scan response (FIG. 2B),and CT scan data (FIG. 2C) for Patient 2.

FIGS. 3A-B show the bone scan (FIG. 3A), bone scan response (FIG. 3B)for Patient 3.

DETAILED DESCRIPTION OF THE INVENTION Abbreviations and Definitions

The following abbreviations and terms have the indicated meaningsthroughout:

Abbreviation Meaning Ac Acetyl bALP Bone-specific alkaline phosphataseBr Broad ° C. Degrees Celsius c- Cyclo CBZ CarboBenZoxy =benzyloxycarbonyl CTx Cross-linked C-terminal telopeptides of type-1collagen d Doublet dd Doublet of doublet dt Doublet of triplet DCMDichloromethane DME 1,2-dimethoxyethane DMF N,N-Dimethylformamide DMSOdimethyl sulfoxide Dppf 1,1’-bis(diphenylphosphano)ferrocene EI ElectronImpact ionization G Gram(s) h or hr Hour(s) HPLC High pressure liquidchromatography L Liter(s) M Molar or molarity m Multiplet MgMilligram(s) MHz Megahertz (frequency) Min Minute(s) mL Milliliter(s) μLMicroliter(s) μM Micromole(s) or micromolar mM Millimolar MmolMillimole(s) Mol Mole(s) MS Mass spectral analysis N Normal or normalitynM Nanomolar NMR Nuclear magnetic resonance spectroscopy NTxCross-linked N-terminal telopeptides of type-1 collagen q Quartet RTRoom temperature s Singlet t or tr Triplet TFA Trifluoroacetic acid THFTetrahydrofuran TLC Thin layer chromatography

The symbol “—” means a single bond, “═” means a double bond.

When chemical structures are depicted or described, unless explicitlystated otherwise, all carbons are assumed to have hydrogen substitutionto conform to a valence of four. For example, in the structure on theleft-hand side of the schematic below there are nine hydrogens implied.The nine hydrogens are depicted in the right-hand structure. Sometimes aparticular atom in a structure is described in textual formula as havinga hydrogen or hydrogens as substitution (expressly defined hydrogen),for example, —CH₂CH₂—. It is understood by one of ordinary skill in theart that the aforementioned descriptive techniques are common in thechemical arts to provide brevity and simplicity to description ofotherwise complex structures.

If a group “R” is depicted as “floating” on a ring system, as forexample in the formula:

then, unless otherwise defined, a substituent “R” may reside on any atomof the ring system, assuming replacement of a depicted, implied, orexpressly defined hydrogen from one of the ring atoms, so long as astable structure is formed.

If a group “R” is depicted as floating on a fused ring system, as forexample in the formulae:

then, unless otherwise defined, a substituent “R” may reside on any atomof the fused ring system, assuming replacement of a depicted hydrogen(for example the —NH— in the formula above), implied hydrogen (forexample as in the formula above, where the hydrogens are not shown butunderstood to be present), or expressly defined hydrogen (for examplewhere in the formula above, “Z” equals=CH—) from one of the ring atoms,so long as a stable structure is formed. In the example depicted, the“R” group may reside on either the 5-membered or the 6-membered ring ofthe fused ring system. When a group “R” is depicted as existing on aring system containing saturated carbons, as for example in the formula:

where, in this example, “y” can be more than one, assuming each replacesa currently depicted, implied, or expressly defined hydrogen on thering; then, unless otherwise defined, where the resulting structure isstable, two “R's” may reside on the same carbon. A simple example iswhen R is a methyl group; there can exist a geminal dimethyl on a carbonof the depicted ring (an “annular” carbon). In another example, two R'son the same carbon, including that carbon, may form a ring, thuscreating a spirocyclic ring (a “spirocyclyl” group) structure with thedepicted ring as for example in the formula:

“Halogen” or “halo” refers to fluorine, chlorine, bromine or iodine.

“Yield” for each of the reactions described herein is expressed as apercentage of the theoretical yield.

“Patient” for the purposes of the present invention includes humans andother animals, particularly mammals, and other organisms. Thus themethods are applicable to both human therapy and veterinaryapplications. In another embodiment the patient is a mammal, and inanother embodiment the patient is human.

A “pharmaceutically acceptable salt” of a compound means a salt that ispharmaceutically acceptable and that possesses the desiredpharmacological activity of the parent compound. It is understood thatthe pharmaceutically acceptable salts are non-toxic. Additionalinformation on suitable pharmaceutically acceptable salts can be foundin Remington's Pharmaceutical Sciences, 17^(th) ed., Mack PublishingCompany, Easton, Pa., 1985, which is incorporated herein by reference orS. M. Berge, et al., “Pharmaceutical Salts,” J. Pharm. Sci., 1977;66:1-19 both of which are incorporated herein by reference.

Examples of pharmaceutically acceptable acid addition salts includethose formed with inorganic acids such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid, and the like; as wellas organic acids such as acetic acid, trifluoroacetic acid, propionicacid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvicacid, lactic acid, oxalic acid, maleic acid, malonic acid, succinicacid, fumaric acid, tartaric acid, malic acid, citric acid, benzoicacid, cinnamic acid, 3-(4-hydroxybenzoyl)benzoic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid, glucoheptonic acid,4,4′-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionicacid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuricacid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylicacid, stearic acid, muconic acid, p-toluenesulfonic acid, and salicylicacid and the like.

“Prodrug” refers to compounds that are transformed (typically rapidly)in vivo to yield the parent compound of the above formulae, for example,by hydrolysis in blood. Common examples include, but are not limited to,ester and amide forms of a compound having an active form bearing acarboxylic acid moiety. Examples of pharmaceutically acceptable estersof the compounds of this invention include, but are not limited to,alkyl esters (for example with between about one and about six carbons)the alkyl group is a straight or branched chain. Acceptable esters alsoinclude cycloalkyl esters and arylalkyl esters such as, but not limitedto benzyl. Examples of pharmaceutically acceptable amides of thecompounds of this invention include, but are not limited to, primaryamides, and secondary and tertiary alkyl amides (for example withbetween about one and about six carbons). Amides and esters of thecompounds of the present invention may be prepared according toconventional methods. A thorough discussion of prodrugs is provided inT. Higuchi and V. Stella, “Pro-drugs as Novel Delivery Systems,” Vol 14of the A.C.S. Symposium Series, and in Bioreversible Carriers in DrugDesign, ed. Edward B. Roche, American Pharmaceutical Association andPergamon Press, 1987, both of which are incorporated herein by referencefor all purposes.

“Therapeutically effective amount” is an amount of a compound of theinvention, that when administered to a patient, ameliorates a symptom ofthe disease. A therapeutically effective amount is intended to includean amount of a compound alone or in combination with other activeingredients effective to modulate c-Met, and/or VEGFR2, or effective totreat or prevent cancer. The amount of a compound of the invention whichconstitutes a “therapeutically effective amount” will vary depending onthe compound, the disease state and its severity, the age of the patientto be treated, and the like. The therapeutically effective amount can bedetermined by one of ordinary skill in the art having regard to theirknowledge and to this disclosure.

“Treating” or “treatment” of a disease, disorder, or syndrome, as usedherein, includes (i) preventing the disease, disorder, or syndrome fromoccurring in a human, i.e. causing the clinical symptoms of the disease,disorder, or syndrome not to develop in an animal that may be exposed toor predisposed to the disease, disorder, or syndrome but does not yetexperience or display symptoms of the disease, disorder, or syndrome;(ii) inhibiting the disease, disorder, or syndrome, i.e., arresting itsdevelopment; and (iii) relieving the disease, disorder, or syndrome,i.e., causing regression of the disease, disorder, or syndrome. As isknown in the art, adjustments for systemic versus localized delivery,age, body weight, general health, sex, diet, time of administration,drug interaction and the severity of the condition may be necessary, andwill be ascertainable with routine experience.

EMBODIMENTS

In one embodiment the compound of Formula I is the compound of FormulaIa:

or a pharmaceutically acceptable salt thereof, wherein:

R¹ is halo;

R² is halo; and

Q is CH or N.

In another embodiment, the compound of Formula I is Compound 1:

or a pharmaceutically acceptable salt thereof. As indicated previously,compound 1 is referred to herein asN-(4-{[6,7-bis(methyloxy)quinolin-4-yl]oxy}phenyl)-N′-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide.WO 2005/030140 discloses Compound 1 and describes how it is made(Example 37, 38, and 48) and also discloses the therapeutic activity ofthis compound to inhibit, regulate and/or modulate the signaltransduction of kinases, (Assays, Table 4, entry 289). Example 48 is onparagraph [0353] in WO 2005/030140, the entire contents of which isincorporated herein by reference.

In other embodiments, the compound of Formula I, Ia, or Compound 1, or apharmaceutically acceptable salt thereof, is administered as apharmaceutical composition, wherein the pharmaceutical compositionadditionally comprises a pharmaceutically acceptable carrier, excipient,or diluent. In a specific embodiment, the Compound of Formula I isCompound 1.

The compound of Formula I, Formula Ia and Compound I, as describedherein, includes both the recited compounds as well as individualisomers and mixtures of isomers. In each instance, the compound ofFormula I includes the pharmaceutically acceptable salts, hydrates,and/or solvates of the recited compounds and any individual isomers ormixture of isomers thereof.

In other embodiments, the compound of Formula I, Ia, or Compound 1 canbe the (L)-malate salt. The malate salt of the Compound of Formula I andof Compound 1 is disclosed in WO 2010/083414 and US 2012/0070368, theentire contents of each of which are incorporated herein by reference.

In other embodiments, the compound of Formula I can be the malate salt.

In other embodiments, the compound of Formula I can be the (D)-malatesalt.

In other embodiments, the compound of Formula I can be the (L)-malatesalt.

In other embodiments, the compound of Formula Ia can be the malate salt.

In other embodiments, the compound of Formula Ia can be the (D)-malatesalt.

In other embodiments, the compound of Formula Ia can be the (L)-malatesalt.

In other embodiments, Compound 1 can be the malate salt.

In other embodiments, Compound 1 can be the (D)-malate salt.

In other embodiments, Compound 1 can be the (L)-malate salt.

In another embodiment, the malate salt is in the crystalline N-1 form ofthe (L) malate salt and/or the (D) malate salt of the Compound 1 asdisclosed in US 2012/0070368, the entire contents of which isincorporated herein by reference. Also see WO 2008/083319, the entirecontents of which is incorporated herein by reference, for theproperties of crystalline enantiomers, including the N-1 and/or the N-2crystalline forms of the malate salt of Compound 1. Methods of makingand characterizing such forms are fully described in WO 2010/083414,which is incorporated herein by reference in its entirety.

In another embodiment, the invention is directed to a method forameliorating the symptoms of osteolytic bone metastases, comprisingadministering to a patient in need of such treatment a therapeuticallyeffective amount of a compound of Formula I in any of the embodimentsdisclosed herein. In a specific embodiment, the Compound of Formula I isCompound 1.

In another embodiment, the compound of Formula I is administeredpost-taxotere treatment. In a specific embodiment, the Compound ofFormula I is Compound 1.

In another embodiment, the compound of Formula I is as effective or moreeffective than mitoxantrone plus prednisone. In a specific embodiment,the Compound of Formula I is Compound 1.

In another embodiment, the Compound of Formula I, Ia, or Compound 1 or apharmaceutically acceptable salt thereof is administered orally oncedaily as a tablet or capsule.

In another embodiment, Compound 1 is administered orally as its freebase or malate salt as a capsule or tablet.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing upto 100 mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing100 mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 95mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 90mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 85mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 80mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 75mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 70mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 65mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 60mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 55mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 50mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 45mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 40mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 30mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 25mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 20mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 15mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 10mg of Compound 1.

In another embodiment, Compound 1 is administered orally once daily asits free base or as the malate salt as a capsule or tablet containing 5mg of Compound 1.

In another embodiment, Compound 1 is administered as its free base ormalate salt orally once daily as a tablet as provided in the followingtable.

Ingredient (% w/w) Compound 1 31.68 Microcrystalline Cellulose 38.85Lactose anhydrous 19.42 Hydroxypropyl Cellulose 3.00 CroscarmelloseSodium 3.00 Total Intra-granular 95.95 Silicon dioxide, Colloidal 0.30Croscarmellose Sodium 3.00 Magnesium Stearate 0.75 Total 100.00

In another embodiment, Compound 1 is administered orally as its freebase or malate salt once daily as a tablet as provided in the followingtable.

Ingredient (% w/w) Compound 1 25.0-33.3 Microcrystalline Cellulose q.sHydroxypropyl Cellulose 3 Poloxamer 0-3 Croscarmellose Sodium 6.0Colloidal Silicon Dioxide 0.5 Magnesium Stearate 0.5-1.0 Total 100

In another embodiment, Compound 1 is administered orally as its freebase or malate salt once daily as a tablet as provided in the followingtable.

Theoretical Quantity Ingredient (mg/unit dose) Compound 1 100.0Microcrystalline Cellulose 155.4 PH-102 Lactose Anhydrous 60M 77.7Hydroxypropyl Cellulose, EXF 12.0 Croscarmellose Sodium 24 ColloidalSilicon Dioxide 1.2 Magnesium Stearate (Non- 3.0 Bovine) Opadry Yellow16.0 Total 416

Any of the tablet formulations provided above can be adjusted accordingto the dose of Compound 1 desired. Thus, the amount of each of theformulation ingredients can be proportionally adjusted to provide atable formulation containing various amounts of Compound 1 as providedin the previous paragraphs. In another embodiment, the formulations cancontain 20, 40, 60, or 80 mg of Compound 1.

Administration

Administration of the compound of Formula I, Formula Ia, or Compound 1,or a pharmaceutically acceptable salt thereof, in pure form or in anappropriate pharmaceutical composition, can be carried out via any ofthe accepted modes of administration or agents for serving similarutilities. Thus, administration can be, for example, orally, nasally,parenterally (intravenous, intramuscular, or subcutaneous), topically,transdermally, intravaginally, intravesically, intracistemally, orrectally, in the form of solid, semi-solid, lyophilized powder, orliquid dosage forms, such as for example, tablets, suppositories, pills,soft elastic and hard gelatin dosages (which can be in capsules ortablets), powders, solutions, suspensions, or aerosols, or the like,specifically in unit dosage forms suitable for simple administration ofprecise dosages.

The compositions will include a conventional pharmaceutical carrier orexcipient and a compound of Formula I as the/an active agent, and, inaddition, may include carriers and adjuvants, etc.

Adjuvants include preserving, wetting, suspending, sweetening,flavoring, perfuming, emulsifying, and dispensing agents. Prevention ofthe action of microorganisms can be ensured by various antibacterial andantifungal agents, for example, parabens, chlorobutanol, phenol, sorbicacid, and the like. It may also be desirable to include isotonic agents,for example sugars, sodium chloride, and the like. Prolonged absorptionof the injectable pharmaceutical form can be brought about by the use ofagents delaying absorption, for example, aluminum monostearate andgelatin.

If desired, a pharmaceutical composition of the compound of Formula Imay also contain minor amounts of auxiliary substances such as wettingor emulsifying agents, pH buffering agents, antioxidants, and the like,such as, for example, citric acid, sorbitan monolaurate, triethanolamineoleate, butylalted hydroxytoluene, etc.

The choice of composition depends on various factors such as the mode ofdrug administration (e.g., for oral administration, compositions in theform of tablets, pills or capsules) and the bioavailability of the drugsubstance. Recently, pharmaceutical compositions have been developedespecially for drugs that show poor bioavailability based upon theprinciple that bioavailability can be increased by increasing thesurface area i.e., decreasing particle size. For example, U.S. Pat. No.4,107,288 describes a pharmaceutical composition having particles in thesize range from 10 to 1,000 nm in which the active material is supportedon a crosslinked matrix of macromolecules. U.S. Pat. No. 5,145,684describes the production of a pharmaceutical composition in which thedrug substance is pulverized to nanoparticles (average particle size of400 nm) in the presence of a surface modifier and then dispersed in aliquid medium to give a pharmaceutical composition that exhibitsremarkably high bioavailability.

Compositions suitable for parenteral injection may comprisephysiologically acceptable sterile aqueous or nonaqueous solutions,dispersions, suspensions or emulsions, and sterile powders forreconstitution into sterile injectable solutions or dispersions.Examples of suitable aqueous and nonaqueous carriers, diluents, solventsor vehicles include water, ethanol, polyols (propyleneglycol,polyethyleneglycol, glycerol, and the like), suitable mixtures thereof,vegetable oils (such as olive oil) and injectable organic esters such asethyl oleate. Proper fluidity can be maintained, for example, by the useof a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersions and by the use of surfactants.

One specific route of administration is oral, using a convenient dailydosage regimen that can be adjusted according to the degree of severityof the disease-state to be treated.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is admixed with at least one inert customary excipient (orcarrier) such as sodium citrate or dicalcium phosphate or (a) fillers orextenders, as for example, starches, lactose, sucrose, glucose,mannitol, and silicic acid, (b) binders, as for example, cellulosederivatives, starch, alignates, gelatin, polyvinylpyrrolidone, sucrose,and gum acacia, (c) humectants, as for example, glycerol, (d)disintegrating agents, as for example, agar-agar, calcium carbonate,potato or tapioca starch, alginic acid, croscarmellose sodium, complexsilicates, and sodium carbonate, (e) solution retarders, as for exampleparaffin, (f) absorption accelerators, as for example, quaternaryammonium compounds, (g) wetting agents, as for example, cetyl alcohol,and glycerol monostearate, magnesium stearate and the like (h)adsorbents, as for example, kaolin and bentonite, and (i) lubricants, asfor example, talc, calcium stearate, magnesium stearate, solidpolyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In thecase of capsules, tablets, and pills, the dosage forms may also comprisebuffering agents.

Solid dosage forms as described above can be prepared with coatings andshells, such as enteric coatings and others well known in the art. Theymay contain pacifying agents, and can also be of such composition thatthey release the active compound or compounds in a certain part of theintestinal tract in a delayed manner. Examples of embedded compositionsthat can be used are polymeric substances and waxes. The activecompounds can also be in microencapsulated form, if appropriate, withone or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, and elixirs. Suchdosage forms are prepared, for example, by dissolving, dispersing, etc.,the compound of Formula I, or a pharmaceutically acceptable saltthereof, and optional pharmaceutical adjuvants in a carrier, such as,for example, water, saline, aqueous dextrose, glycerol, ethanol and thelike; solubilizing agents and emulsifiers, as for example, ethylalcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzylalcohol, benzyl benzoate, propyleneglycol, 1,3-butyleneglycol,dimethylformamide; oils, in particular, cottonseed oil, groundnut oil,corn germ oil, olive oil, castor oil and sesame oil, glycerol,tetrahydrofurfuryl alcohol, polyethyleneglycols and fatty acid esters ofsorbitan; or mixtures of these substances, and the like, to thereby forma solution or suspension.

Suspensions, in addition to the active compounds, may contain suspendingagents, as for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, or mixtures of thesesubstances, and the like.

Compositions for rectal administration are, for example, suppositoriesthat can be prepared by mixing the compound of Formula I with, forexample, suitable non-irritating excipients or carriers such as cocoabutter, polyethyleneglycol or a suppository wax, which are solid atordinary temperatures but liquid at body temperature and therefore, meltwhile in a suitable body cavity and release the active componenttherein.

Dosage forms for topical administration of the compound of Formula Iinclude ointments, powders, sprays, and inhalants. The active componentis admixed under sterile conditions with a physiologically acceptablecarrier and any preservatives, buffers, or propellants as may berequired. Ophthalmic compositions, eye ointments, powders, and solutionsare also contemplated as being within the scope of this disclosure.

Compressed gases may be used to disperse the compound of Formula I inaerosol form. Inert gases suitable for this purpose are nitrogen, carbondioxide, etc.

Generally, depending on the intended mode of administration, thepharmaceutically acceptable compositions will contain about 1% to about99% by weight of a compound(s) of Formula I, or a pharmaceuticallyacceptable salt thereof, and 99% to 1% by weight of a suitablepharmaceutical excipient. In one example, the composition will bebetween about 5% and about 75% by weight of a compound(s) of Formula I,Formula Ia, or Compound 1, or a pharmaceutically acceptable saltthereof, with the rest being suitable pharmaceutical excipients.

Actual methods of preparing such dosage forms are known, or will beapparent, to those skilled in this art; for example, see Remington'sPharmaceutical Sciences, 18th Ed., (Mack Publishing Company, Easton,Pa., 1990). The composition to be administered will, in any event,contain a therapeutically effective amount of a compound of Formula I,or a pharmaceutically acceptable salt thereof, for treatment of adisease-state in accordance with the teachings of this disclosure.

The compounds of this disclosure, or their pharmaceutically acceptablesalts or solvates, are administered in a therapeutically effectiveamount which will vary depending upon a variety of factors including theactivity of the specific compound employed, the metabolic stability andlength of action of the compound, the age, body weight, general health,sex, diet, mode and time of administration, rate of excretion, drugcombination, the severity of the particular disease-states, and the hostundergoing therapy. The compound of Formula I, Formula Ia, or Compound1, can be administered to a patient at dosage levels in the range ofabout 0.1 to about 1,000 mg per day. For a normal human adult having abody weight of about 70 kilograms, a dosage in the range of about 0.01to about 100 mg per kilogram of body weight per day is an example. Thespecific dosage used, however, can vary. For example, the dosage candepend on a number of factors including the requirements of the patient,the severity of the condition being treated, and the pharmacologicalactivity of the compound being used. The determination of optimumdosages for a particular patient is well known to one of ordinary skillin the art.

In other embodiments, the compound of Formula I, Formula Ia, or Compound1, can be administered to the patient concurrently with other cancertreatments. Such treatments include other cancer chemotherapeutics,hormone replacement therapy, radiation therapy, or immunotherapy, amongothers. The choice of other therapy will depend on a number of factorsincluding the metabolic stability and length of action of the compound,the age, body weight, general health, sex, diet, mode and time ofadministration, rate of excretion, drug combination, the severity of theparticular disease-states, and the host undergoing therapy.

Preparation of Compound 1

Preparation ofN-(4-{[6,7-bis(methyloxy)quinolin-4-yl]oxy}phenyl)-N′-(4-fluorophenyl)cyclopropane-1,1-dicarboxamideand the (L)-malate salt thereof.

The synthetic route used for the preparation ofN-(4-[6,7-bis(methyloxy)quinolin-4-yl]oxy}phenyl)-N′-(4-fluorophenyl)cyclopropane-1,1-dicarboxamideand the (L)-malate salt thereof is depicted in Scheme 1:

Preparation of 4-Chloro-6,7-dimethoxy-quinoline

A reactor was charged sequentially with 6,7-dimethoxy-quinoline-4-ol(10.0 kg) and acetonitrile (64.0 L). The resulting mixture was heated toapproximately 65° C. and phosphorus oxychloride (POCl₃, 50.0 kg) wasadded. After the addition of POCl₃, the temperature of the reactionmixture was raised to approximately 80° C. The reaction was deemedcomplete (approximately 9.0 hours) when less than 2 percent of thestarting material remained (in process high-performance liquidchromotography [HPLC] analysis). The reaction mixture was cooled toapproximately 10° C. and then quenched into a chilled solution ofdichloromethane (DCM, 238.0 kg), 30% NH₄OH (135.0 kg), and ice (440.0kg). The resulting mixture was warmed to approximately 14° C., andphases were separated. The organic phase was washed with water (40.0 kg)and concentrated by vacuum distillation to remove the solvent(approximately 190.0 kg). Methyl-t-butyl ether (MTBE, 50.0 kg) was addedto the batch, and the mixture was cooled to approximately 10° C., duringwhich time the product crystallized out. The solids were recovered bycentrifugation, washed with n heptane (20.0 kg), and dried atapproximately 40° C. to afford the title compound (8.0 kg).

Preparation of 6,7-Dimethyl-4-(4-nitro-phenoxy)-quinoline

A reactor was sequentially charged with 4-chloro-6,7-dimethoxy-quinoline(8.0 kg), 4 nitrophenol (7.0 kg), 4 dimethylaminopyridine (0.9 kg), and2,6 lutidine (40.0 kg). The reactor contents were heated toapproximately 147° C. When the reaction was complete (less than 5percent starting material remaining as determined by in process HPLCanalysis, approximately 20 hours), the reactor contents were allowed tocool to approximately 25° C. Methanol (26.0 kg) was added, followed bypotassium carbonate (3.0 kg) dissolved in water (50.0 kg). The reactorcontents were stirred for approximately 2 hours. The resulting solidprecipitate was filtered, washed with water (67.0 kg), and dried at 25°C. for approximately 12 hours to afford the title compound (4.0 kg).

Preparation of 4-(6,7-Dimethoxy-quinoline-4-yloxy)-phenylamine

A solution containing potassium formate (5.0 kg), formic acid (3.0 kg),and water (16.0 kg) was added to a mixture of6,7-dimethoxy-4-(4-nitro-phenoxy)-quinoline (4.0 kg), 10 percentpalladium on carbon (50 percent water wet, 0.4 kg) in tetrahydrofuran(THF, 40.0 kg) that had been heated to approximately 60° C. The additionwas carried out such that the temperature of the reaction mixtureremained approximately 60° C. When the reaction was deemed complete asdetermined using in-process HPLC analysis (less than 2 percent startingmaterial remaining, typically 15 hours), the reactor contents werefiltered. The filtrate was concentrated by vacuum distillation atapproximately 35° C. to half of its original volume, which resulted inthe precipitation of the product. The product was recovered byfiltration, washed with water (12.0 kg), and dried under vacuum atapproximately 50° C. to afford the title compound (3.0 kg; 97 percentarea under curve (AUC)).

Preparation of 1-(4-Fluoro-phenylcarbamoyl)-cyclopropanecarboxylic acid

Triethylamine (8.0 kg) was added to a cooled (approximately 4° C.)solution of commercially available cyclopropane-1,1-dicarboxylic acid (2l, 10.0 kg) in THF (63.0 kg) at a rate such that the batch temperaturedid not exceed 10° C. The solution was stirred for approximately 30minutes, and then thionyl chloride (9.0 kg) was added, keeping the batchtemperature below 10° C. When the addition was complete, a solution of4-fluoroaniline (9.0 kg) in THF (25.0 kg) was added at a rate such thatthe batch temperature did not exceed 10° C. The mixture was stirred forapproximately 4 hours and then diluted with isopropyl acetate (87.0 kg).This solution was washed sequentially with aqueous sodium hydroxide (2.0kg dissolved in 50.0 L of water), water (40.0 L), and aqueous sodiumchloride (10.0 kg dissolved in 40.0 L of water). The organic solutionwas concentrated by vacuum distillation followed by the addition ofheptane, which resulted in the precipitation of solid. The solid wasrecovered by centrifugation and then dried at approximately 35° C. undervacuum to afford the title compound. (10.0 kg).

Preparation of 1-(4-Fluoro-phenylcarbamoyl)-cyclopropanecarbonylchloride

Oxalyl chloride (1.0 kg) was added to a solution of1-(4-fluoro-phenylcarbamoyl)-cyclopropanecarboxylic acid (2.0 kg) in amixture of THF (11 kg) and N, N-dimethylformamide (DMF; 0.02 kg) at arate such that the batch temperature did not exceed 30° C. This solutionwas used in the next step without further processing.

Preparation ofN-(4-{[6,7-bis(methyloxy)quinolin-4-yl]oxy}phenyl)-N′-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide

The solution from the previous step containing1-(4-fluoro-phenylcarbamoyl)-cyclopropanecarbonyl chloride was added toa mixture of 4-(6,7-dimethoxy-quinoline-4-yloxy)-phenylamine (3.0 kg)and potassium carbonate (4.0 kg) in THF (27.0 kg) and water (13.0 kg) ata rate such that the batch temperature did not exceed 30° C. When thereaction was complete (in typically 10 minutes), water (74.0 kg) wasadded. The mixture was stirred at 15-30° C. for approximately 10 hours,which resulted in the precipitation of the product. The product wasrecovered by filtration, washed with a pre-made solution of THF (11.0kg) and water (24.0 kg), and dried at approximately 65° C. under vacuumfor approximately 12 hours to afford the title compound (free base, 5.0kg). ¹H NMR (400 MHz, d₆-DMSO): δ 10.2 (s, 1H), 10.05 (s, 1H), 8.4 (s,1H), 7.8 (m, 2H), 7.65 (m, 2H), 7.5 (s, 1H), 7.35 (s, 1H), 7.25 (m, 2H),7.15 (m, 2H), 6.4 (s, 1H), 4.0 (d, 6H), 1.5 (s, 4H). LC/MS: M+H=502.

Preparation ofN-(4-{[6,7-bis(methyloxy)quinolin-4-yl]oxy}phenyl)-N′-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide,(L) malate salt

A solution of L-malic acid (2.0 kg) in water (2.0 kg) was added to asolution of Cyclopropane-1,1-dicarboxylic acid[4-(6,7-dimethoxy-quinoline-4-yloxy)-phenyl]-amide(4-fluoro-phenyl)-amide free base (15, 5.0 kg) in ethanol, maintaining abatch temperature of approximately 25° C. Carbon (0.5 kg) and thiolsilica (0.1 kg) were then added, and the resulting mixture was heated toapproximately 78° C., at which point water (6.0 kg) was added. Thereaction mixture was then filtered, followed by the addition ofisopropanol (38.0 kg), and was allowed to cool to approximately 25° C.The product was recovered by filtration and washed with isopropanol(20.0 kg), and dried at approximately 65° C. to afford the titlecompound (5.0 kg).

Alternative Preparation ofN-(4-{[6,7-Bis(methyloxy)quinolin-4-yl]oxy}phenyl)-N′-(4-fluorophenyl)cyclopropane-1,1-dicarboxamideand the (L)-malate salt thereof

An alternative synthetic route that can be used for the preparation ofN-(4-{[6,7-bis(methyloxy)quinolin-4-yl]oxy}phenyl)-N′-(4-fluorophenyl)cyclopropane-1,1-dicarboxamideand the (L)-malate salt thereof is depicted in Scheme 2.

Preparation of 4-Chloro-6,7-dimethoxy-quinoline

A reactor was charged sequentially with 6,7-dimethoxy-quinoline-4-ol(47.0 kg) and acetonitrile (318.8 kg). The resulting mixture was heatedto approximately 60° C. and phosphorus oxychloride (POCl₃, 130.6 kg) wasadded. After the addition of POCl₃, the temperature of the reactionmixture was raised to approximately 77° C. The reaction was deemedcomplete (approximately 13 hours) when less than 3% of the startingmaterial remained (in-process high-performance liquid chromatography[HPLC] analysis). The reaction mixture was cooled to approximately 2-7°C. and then quenched into a chilled solution of dichloromethane (DCM,482.8 kg), 26 percent NH₄OH (251.3 kg), and water (900 L). The resultingmixture was warmed to approximately 20-25° C., and phases wereseparated. The organic phase was filtered through a bed of AW hyflosuper-cel NF (Celite; 5.4 kg) and the filter bed was washed with DCM(118.9 kg). The combined organic phase was washed with brine (282.9 kg)and mixed with water (120 L). The phases were separated and the organicphase was concentrated by vacuum distillation with the removal ofsolvent (approximately 95 L residual volume). DCM (686.5 kg) was chargedto the reactor containing organic phase and concentrated by vacuumdistillation with the removal of solvent (approximately 90 L residualvolume). Methyl t-butyl ether (MTBE, 226.0 kg) was then charged and thetemperature of the mixture was adjusted to −20 to −25° C. and held for2.5 hours resulting in solid precipitate which was then filtered andwashed with n-heptane (92.0 kg), and dried on a filter at approximately25° C. under nitrogen to afford the title compound. (35.6 kg).

Preparation of 4-(6,7-Dimethoxy-quinoline-4-yloxy)-phenylamine

4-Aminophenol (24.4 kg) dissolved in N,N-dimethylacetamide (DMA, 184.3kg) was charged to a reactor containing 4-chloro-6,7-dimethoxyquinoline(35.3 kg), sodium t-butoxide (21.4 kg) and DMA (167.2 kg) at 20-25° C.This mixture was then heated to 100-105° C. for approximately 13 hours.After the reaction was deemed complete as determined using in-processHPLC analysis (less than 2 percent starting material remaining), thereactor contents were cooled at 15-20° C. and water (pre-cooled, 2-7°C., 587 L) charged at a rate to maintain 15-30° C. temperature. Theresulting solid precipitate was filtered, washed with a mixture of water(47 L) and DMA (89.1 kg) and finally with water (214 L). The filter cakewas then dried at approximately 25° C. on filter to yield crude4-(6,7-dimethoxy-quinoline-4-yloxy)-phenylamine (59.4 kg wet, 41.6 kgdry calculated based on LOD). Crude4-(6,7-dimethoxy-quinoline-4-yloxy)-phenylamine was refluxed(approximately 75° C.) in a mixture of tetrahydrofuran (THF, 211.4 kg)and DMA (108.8 kg) for approximately 1 hour and then cooled to 0-5° C.and aged for approximately 1 hour after which time the solid wasfiltered, washed with THF (147.6 kg) and dried on a filter under vacuumat approximately 25° C. to yield4-(6,7-dimethoxy-quinoline-4-yloxy)-phenylamine (34.0 kg).

Alternative Preparation of4-(6,7-Dimethoxy-quinoline-4-yloxy)-phenylamine

4-chloro-6,7-dimethoxyquinoline (34.8 kg) and 4-aminophenol (30.8 kg)and sodium tert pentoxide (1.8 equivalents) 88.7 kg, 35 weight percentin THF) were charged to a reactor, followed by N,N-dimethylacetamide(DMA, 293.3 kg). This mixture was then heated to 105-115° C. forapproximately 9 hours. After the reaction was deemed complete asdetermined using in-process HPLC analysis (less than 2 percent startingmaterial remaining), the reactor contents were cooled at 15-25° C. andwater (315 kg) was added over a two hour period while maintaining thetemperature between 20-30° C. The reaction mixture was then agitated foran additional hour at 20-25° C. The crude product was collected byfiltration and washed with a mixture of 88 kg water and 82.1 kg DMA,followed by 175 kg water. The product was dried on a filter drier for 53hours. The LOD showed less than 1 percent w/w.

In an alternative procedure, 1.6 equivalents of sodium tert-pentoxidewere used and the reaction temperature was increased from 110-120° C. Inaddition, the cool down temperature was increased to 35-40° C. and thestarting temperature of the water addition was adjusted to 35-40° C.,with an allowed exotherm to 45° C.

Preparation of 1-(4-Fluorophenylcarbamoyl)-cyclopropanecarboxylic acid

Triethylamine (19.5 kg) was added to a cooled (approximately 5° C.)solution of cyclopropane-1,1-dicarboxylic acid (24.7 kg) in THF (89.6kg) at a rate such that the batch temperature did not exceed 5° C. Thesolution was stirred for approximately 1.3 hours, and then thionylchloride (23.1 kg) was added, keeping the batch temperature below 10° C.When the addition was complete, the solution was stirred forapproximately 4 hours keeping temperature below 10° C. A solution of4-fluoroaniline (18.0 kg) in THF (33.1 kg) was then added at a rate suchthat the batch temperature did not exceed 10° C. The mixture was stirredfor approximately 10 hours after which the reaction was deemed complete.The reaction mixture was then diluted with isopropyl acetate (218.1 kg).This solution was washed sequentially with aqueous sodium hydroxide(10.4 kg, 50 percent dissolved in 119 L of water) further diluted withwater (415 L), then with water (100 L) and finally with aqueous sodiumchloride (20.0 kg dissolved in 100 L of water). The organic solution wasconcentrated by vacuum distillation (100 L residual volume) below 40° C.followed by the addition of n-heptane (171.4 kg), which resulted in theprecipitation of solid. The solid was recovered by filtration and washedwith n-heptane (102.4 kg), resulting in wet, crude1-(4-fluoro-phenylcarbamoyl)-cyclopropanecarboxylic acid (29.0 kg). Thecrude, 1-(4-fluoro-phenylcarbamoyl)-cyclopropanecarboxylic acid wasdissolved in methanol (139.7 kg) at approximately 25° C. followed by theaddition of water (320 L) resulting in slurry which was recovered byfiltration, washed sequentially with water (20 L) and n-heptane (103.1kg) and then dried on the filter at approximately 25° C. under nitrogento afford the title compound (25.4 kg).

Preparation of 1-(4-Fluoro-phenylcarbamoyl)-cyclopropanecarbonylchloride

Oxalyl chloride (12.6 kg) was added to a solution of1-(4-fluoro-phenylcarbamoyl)-cyclopropanecarboxylic acid (22.8 kg) in amixture of THF (96.1 kg) and N, N-dimethylformamide (DMF; 0.23 kg) at arate such that the batch temperature did not exceed 25° C. This solutionwas used in the next step without further processing.

Alternative Preparation of1-(4-Fluoro-phenylcarbamoyl)-cyclopropanecarbonyl chloride

A reactor was charged with1-(4-fluoro-phenylcarbamoyl)-cyclopropanecarboxylic acid (35 kg), 344 gDMF, and 175 kg THF. The reaction mixture was adjusted to 12-17° C. andthen to the reaction mixture was charged 19.9 kg of oxalyl chloride overa period of 1 hour. The reaction mixture was left stirring at 12-17° C.for 3 to 8 hours. This solution was used in the next step withoutfurther processing.

Preparation of cyclopropane-1,1-dicarboxylic acid[4-(6,7-dimethoxy-quinoline-4-yloxy)-phenyl]-amide(4-fluoro-phenyl)-amide

The solution from the previous step containing1-(4-fluoro-phenylcarbamoyl)-cyclopropanecarbonyl chloride was added toa mixture of compound 4-(6,7-dimethoxy-quinoline-4-yloxy)-phenylamine(23.5 kg) and potassium carbonate (31.9 kg) in THF (245.7 kg) and water(116 L) at a rate such that the batch temperature did not exceed 30° C.When the reaction was complete (in approximately 20 minutes), water (653L) was added. The mixture was stirred at 20-25° C. for approximately 10hours, which resulted in the precipitation of the product. The productwas recovered by filtration, washed with a pre-made solution of THF(68.6 kg) and water (256 L), and dried first on a filter under nitrogenat approximately 25° C. and then at approximately 45° C. under vacuum toafford the title compound (41.0 kg, 38.1 kg, calculated based on LOD).

Alternative Preparation of cyclopropane-1,1-dicarboxylic acid[4-(6,7-dimethoxy-quinoline-4-yloxy)-phenyl]-amide(4-fluoro-phenyl)-amide

A reactor was charged with4-(6,7-dimethoxy-quinoline-4-yloxy)-phenylamine (35.7 kg, 1 equivalent),followed by 412.9 kg THF. To the reaction mixture was charged a solutionof 48.3 K₂CO₃ in 169 kg water. The acid chloride solution of describedin the Alternative Preparation of1-(4-Fluoro-phenylcarbamoyl)-cyclopropanecarbonyl chloride above wastransferred to the reactor containing4-(6,7-dimethoxy-quinoline-4-yloxy)-phenylamine while maintaining thetemperature between 20-30° C. over a minimum of two hours. The reactionmixture was stirred at 20-25° C. for a minimum of three hours. Thereaction temperature was then adjusted to 30-25° C. and the mixture wasagitated. The agitation was stopped and the phases of the mixture wereallowed to separate. The lower aqueous phase was removed and discarded.To the remaining upper organic phase was added 804 kg water. Thereaction was left stirring at 15-25° C. for a minimum of 16 hours.

The product precipitated. The product was filtered and washed with amixture of 179 kg water and 157.9 kg THF in two portions. The crudeproduct was dried under a vacuum for at least two hours. The driedproduct was then taken up in 285.1 kg THF. The resulting suspension wastransferred to reaction vessel and agitated until the suspension becamea clear (dissolved) solution, which required heating to 30-35° C. forapproximately 30 minutes. 456 kg water was then added to the solution,as well as 20 kg SDAG-1 ethanol (ethanol denatured with methanol overtwo hours. The mixture was agitated at 15-25° C. fir at least 16 hours.The product was filtered and washed with a mixture of 143 kg water and126.7 THF in two portions. The product was dried at a maximumtemperature set point of 40° C.

In an alternative procedure, the reaction temperature during acidchloride formation was adjusted to 10-15° C. The recrystallizationtemperature was changed from 15-25° C. to 45-50° C. for 1 hour and thencooled to 15-25° C. over 2 hours.

Preparation of cyclopropane-1,1-dicarboxylic acid[4-(6,7-dimethoxy-quinoline-4-yloxy)-phenyl]-amide(4-fluoro-phenyl)-amide, malate salt

Cyclopropane-1,1-dicarboxylic acid[4-(6,7-dimethoxy-quinoline-4-yloxy)-phenyl]-amide(4-fluoro-phenyl)-amide (1-5; 13.3 kg), L-malic acid (4.96 kg), methylethyl ketone (MEK; 188.6 kg) and water (37.3 kg) were charged to areactor and the mixture was heated to reflux (approximately 74° C.) forapproximately 2 hours. The reactor temperature was reduced to 50 to 55°C. and the reactor contents were filtered. These sequential stepsdescribed above were repeated two more times starting with similaramounts of starting material (13.3 kg), L-Malic acid (4.96 kg), MEK(198.6 kg) and water (37.2 kg). The combined filtrate was azeotropicallydried at atmospheric pressure using MEK (1133.2 kg) (approximateresidual volume 711 L; KF≦0.5% w/w) at approximately 74° C. Thetemperature of the reactor contents was reduced to 20 to 25° C. and heldfor approximately 4 hours resulting in solid precipitate which wasfiltered, washed with MEK (448 kg) and dried under vacuum at 50° C. toafford the title compound (45.5 kg).

Alternative Preparation of cyclopropane-1,1-dicarboxylic acid[4-(6,7-dimethoxy-quinoline-4-yloxy)-phenyl]-amide(4-fluoro-phenyl)-amide, (L) malate salt

Cyclopropane-1,1-dicarboxylic acid[4-(6,7-dimethoxy-quinoline-4-yloxy)-phenyl]-amide(4-fluoro-phenyl)-amide (47.9 kg), L-malic acid (17.2), 658.2 kg methylethyl ketone, and 129.1 kg water (37.3 kg) were charged to a reactor andthe mixture was heated 50-55° C. for approximately 1-3 hours, and thenat 55-60° C. for an addition al 4-5 hours. The mixture was clarified byfiltration through a 1 μm cartridge. The reactor temperature wasadjusted to 20-25° C. and vacuum distilled with a vacuum at 150-200 mmHg with a maximum jacket temperature of 55° C. to the volume range of558-731 L.

The vacuum distillation was performed two more times with the charge of380 kg and 380.2 kg methyl ethyl ketone, respectively. After the thirddistillation, the volume of the batch was adjusted to 18 v/w ofcyclopropane-1,1-dicarboxylic acid[4-(6,7-dimethoxy-quinoline-4-yloxy)-phenyl]-amide(4-fluoro-phenyl)-amide by charging 159.9 kg methyl ethyl ketone to givea total volume of 880 L. An addition al vacuum distillation was carriedout by adjusting 245.7 methyl ethyl ketone. The reaction mixture wasleft with moderate agitation at 20-25° C. for at least 24 hours. Theproduct was filtered and washed with 415.1 kg methyl ethyl ketone inthree portions. The product was dried under a vacuum with the jackettemperature set point at 45° C.

In an alternative procedure, the order of addition was changed so that asolution of 17.7 kg L-malic acid dissolved in 129.9 kg water was addedto cyclopropane-1,1-dicarboxylic acid[4-(6,7-dimethoxy-quinoline-4-yloxy)-phenyl]amide(4-fluoro-phenyl)-amide (48.7 kg) in methyl ethyl ketone (673.3 kg).

Case Studies

The MET and VEGF signaling pathways appear to play important roles inosteoblast and osteoclast function. Strong immunohistochemical stainingof MET has been observed in both cell types in developing bone. HGF andMET are expressed by osteoblasts and osteoclasts in vitro and mediatecellular responses such as proliferation, migration, and expression ofALP. Secretion of HGF by osteoblasts has been proposed as a key factorin osteoblast/osteoclast coupling, and in the development of bonemetastases by tumor cells that express MET. Osteoblasts and osteoclastsalso express VEGF and its receptors, and VEGF signaling in these cellsis involved in potential autocrine and/or paracrine feedback mechanismsregulating cell migration, differentiation, and survival.

Bone metastases are present in 90 percent of patients withcastration-resistant prostate cancer (CRPC), causing significantmorbidity and mortality. Activation of the MET and VEGFR signalingpathways is implicated in the development of bone metastases in CRPC.Three metastatic CRPC patients treated with Compound 1, an inhibitor ofMET and VEGFR, had dramatic responses with near complete resolution ofbone lesions, marked reduction in bone pain and total serum alkalinephosphatase (tALP) levels, and reduction in measurable disease. Theseresults indicate that dual modulation of the MET and VEGFR signalingpathways is a useful therapeutic approach for treating CRPC.

Compound 1 is an orally bioavailable multitargeted tyrosine kinaseinhibitor with potent activity against MET and VEGFR2. Compound 1suppresses MET and VEGFR2 signaling, rapidly induces apoptosis ofendothelial cells and tumor cells, and causes tumor regression inxenograft tumor models. Compound 1 also significantly reduces tumorinvasiveness and metastasis and substantially improves overall survivalin a murine pancreatic neuroendocrine tumor model. In a phase 1 clinicalstudy, Compound 1 was generally well-tolerated, with fatigue, diarrhea,anorexia, rash, and palmar-plantar erythrodysesthesia being the mostcommonly observed adverse events.

Based on target rationale and observed antitumor activity in clinicalstudies, an adaptive phase 2 trial was undertaken in multipleindications including CRPC(http://clinicaltrials.gov/ct2/results?term=NCT00940225 for StudyNCT00940225 last visited Sep. 20, 2011)), in which Compound 1 wasadministered as a 100 mg dose to patients. The findings in the firstthree CRPC patients with evidence of bone metastases on bone scanenrolled to this study are described in the following Case Studies.

Baseline characteristics for patients 1-3 are summarized in Table 1.

TABLE 1 Summary of Baseline Characteristics and Preliminary BestResponses for CRPC Patients Treated with Compound 1. BaselineCharacteristics Patient 1 Patient 2 Patient 3 Age (years) 77 73 66Diagnosis 1993 2009 2009 ECOG performance 1 0 1 status Diseaselocation(s) Lung, Liver, LN, bone LN, bone LN, bone Prior cancer RadicalRadiation to CAB, therapies prostatectomy, pubic ramus docetaxelradiation to and prostate bed, acetabulum, CAB, DES, CAB docetaxelBisphophonates No No Yes Narcotics Yes No No Pain Yes Yes Yes PSA(ng/mL) 430.4 14.7 2.8 tALP (U/L) 689 108 869 Hemoglobin (g/dL) 13.513.3 10.2 Summary of Best Responses Tumor response −41% −20% −51% Bonescan Complete Improvement Near resolution resolution Pain ImprovementPain-free Pain-free PSA −78% +61% −57% tALP −77% −6% −77% Hemoglobin(g/dL) +1.4 +1.8 +2.2 ADT, androgen-deprivation therapy; CAB, combinedandrogen blockade (leuprolide + bicalutamide); DES, diethylstilbestrol;LN, lymph node; PSA, prostate-specific antigen; tALP, total alkalinephosphatase.

Patient 1 was diagnosed with localized prostate cancer in 1993 andtreated with radical prostatectomy (Gleason score unavailable; PSA, 0.99ng/mL). In 2000, local disease recurrence was treated with radiationtherapy. In 2001, combined androgen blockade (CAB) with leuprolide andbicalutamide was initiated for rising PSA (3.5 ng/mL). In 2006,diethystillbestrol (DES) was administered briefly. In 2007, 6 cycles ofdocetaxel were given for new lung metastases. Rising PSA wasunresponsive to antiandrogen withdrawal. Androgen ablation therapy wascontinued until clinical progression. In October 2009, bone metastasisto the spine associated with impingement on the spinal cord and backpain, was treated with radiation therapy (37.5 Gy). In February 2010, abone scan was performed due to increasing bone pain and showed diffuseuptake of radiotracer in the axial and appendicular skeleton. A CT scanrevealed new pulmonary and mediastinal lymph node metastases. PSA was430.4 ng/mL.

Patient 2 was diagnosed in April of 2009 after presenting with apathologic fracture (Gleason score, 4+5=9; PSA, 45.34 ng/mL). Bone scanshowed uptake of radiotracer in the left iliac wing, left sacroiliacjoint, femoral head, and the pubic symphysis. Biopsy of the left pubicramus confirmed metastatic adenocarcinoma with mixed lytic and blasticlesions. CAB with leuprolide and bicalutamide and radiation therapy (8Gy) to the left pubic ramus and acetabulum resulted in bone pain reliefand PSA normalization. Rising PSA in November 2009 (16 ng/mL) wasunresponsive to antiandrogen withdrawal. In February 2010, bone scanshowed multiple foci throughout the axial and appendicular skeleton. ACT scan revealed retroperitoneal lymph node enlargement and livermetastases (PSA, 28.1 ng/mL). Further progression of disease was markedby recurrent bone pain, new lung and hepatic metastases.

Patient 3 was diagnosed in April 2009 after presenting with right hippain (Gleason score, 4+5=9; PSA, 2.6 ng/mL). Bone scan showed uptake ofradiotracer at multiple sites throughout the axial and appendicularskeleton. A CT scan revealed retroperitoneal, common iliac, andsupraclavicular adenopathy. CAB with leuprolide and bicalutamide wasinitiated. The patient received 6 cycles of docetaxel through December2009. Following treatment, a bone scan showed no changes. A CT scanrevealed near resolution of the retroperitoneal and common iliacadenopathy. In March 2010, PSA began to rise, and bone pain worsened. Arepeat bone scan showed new foci, and a CT scan showed an increase inthe retroperitoneal, para-aortic, and bilateral common iliac adenopathy.Rising PSA in April 2010 (2.8 ng/mL) and increasing bone pain wereunresponsive to antiandrogen withdrawal.

Results

All patients provided informed consent before study screening.

Patient 1 started Compound 1 on Feb. 12, 2010. Four weeks later,significant reduction in bone pain was reported. At Week 6, bone scanshowed a dramatic decrease in radiotracer uptake by bone metastases(FIG. 1A). A CT scan showed a partial response (PR) with a 33% decreasein measurable target lesions (FIG. 1C). At Week 12, near completeresolution of bone lesions and a 44% decrease in target lesions wasobserved and was stable through Week 18. Corresponding with the bonescan response, after an initial rise, serum tALP levels decreased from689 U/L at baseline to 159 U/L at Week 18 (FIG. 1B and Table 1). Inaddition, there was an increase in hemoglobin of 1.4 g/dL at Week 2compared with baseline (Table 1). PSA decreased from 430 ng/mL atbaseline to 93.5 ng/mL at Week 18 (FIG. 1B and Table 1). The patient wason open-label treatment through Week 18 when he withdrew afterdeveloping Grade 3 diarrhea.

Patient 2 started Compound 1 on Mar. 31, 2010. At Week 4, reduction inbone pain was reported. At Week 6, bone scan showed a slight flair inradiotracer uptake by bone lesions (FIG. 2A), and a CT scan showed a 13%decrease in target lesions (FIG. 2C). At Week 12, a substantialreduction of radiotracer uptake (FIG. 2A) and a 20% decrease inmeasurable disease were observed (Table 1). After randomization toplacebo at Week 12 the patient developed severe bone pain and sacralnerve root impingement. Radiation to the spine was administered, and thepatient crossed over to open-label Compound 1 treatment at Week 15.Serum tALP levels were within the normal range (101-144 U/L) (FIG. 2B).Hemoglobin increased by 1.8 g/dL at Week 12 compared with baseline(Table 1). PSA peaked at close to 6-fold of baseline by Week 16, butthen decreased to 2-fold of baseline by Week 18 subsequent to crossingover to Compound 1 from placebo (FIG. 2B and Table 1). The patientcontinues on Compound 1 treatment as of September 2010.

Patient 3 started Compound 1 on Apr. 26, 2010. After three weeks acomplete resolution of pain was reported. At Week 6, bone scan showed adramatic reduction in radiotracer uptake (FIG. 3A), and a CT scan showeda PR with a 43% decrease in measurable target lesions. At Week 12 acomplete resolution of bone lesions on bone scan (FIG. 3A) and a 51%decrease in measurable disease were observed (Table 1 and FIG. 3B)).After an initial rise, serum tALP levels steadily decreased, with tALPat 869 U/L at baseline and 197 U/L at Week 18 (FIG. 3B and Table 1).Hemoglobin increased 2.2 g/dL at Week 2 compared with baseline (Table1). PSA decreased from 2.4 ng/mL at screening to 1.2 ng/mL at Week 18(FIG. 3B and Table 1). The patient continues on Compound 1 treatment asof September 2010.

Discussion

All three patients experienced a striking decrease in uptake ofradiotracer on bone scan upon treatment with Compound 1. These findingswere accompanied by substantial reductions in bone pain and evidence ofresponse or stabilization in soft tissue lesions during therapy withCompound 1. The onset of the effect was very rapid in two of thepatients, with substantial improvement or near resolution of bone scanand improvement in pain occurring in the first 6 weeks. In the thirdpatient, an apparent flare in the bone scan was observed at 6 weeks,followed by improvement by 12 weeks. To our knowledge, such acomprehensive and rapid impact on both osseous and soft tissue diseasehas not been observed in this patient population.

Uptake of radiotracer in bone depends on both local blood flow andosteoblastic activity, both of which may be pathologically modulated bythe tumor cells associated with the bone lesion. Resolving uptake maytherefore be attributable to either interruption of local blood flow,direct modulation of osteoblastic activity, a direct effect on the tumorcells in bone, or a combination of these processes. However, decreaseduptake on bone scan in men with CRPC has only been rarely noted withVEGF/VEGFR targeted therapy, despite numerous trials with such agents.Similarly, observations of decreased uptake on bone scan in CRPCpatients have only been reported rarely for abiraterone, which targetsthe cancer cells directly, and for dasatinib, which targets both cancercells and osteoclasts. Thus, targeting angiogenesis alone, orselectively targeting the tumor cells and/or osteoclasts, has notresulted in effects similar to those observed in the patients treatedwith Compound 1.

These results indicate a potential critical role for the MET and VEGFsignaling pathways in the progression of CRPC and point to the promisethat simultaneously targeting these pathways may hold in reducingmorbidity and mortality in this patient population.

Study 2: Effect of Compound 1 Administration on NT^(x) PlasmaConcentration

The effects of Compound 1 treatment on osteoclast activity was alsoinvestigated based on the measurement of changes in plasma concentrationof the bone marker Cross-linked Cross-linked N-terminal telopeptides oftype-1 collagen (NT^(x)) in CRPC patients (N=70).Bisphosphonate/denosumab treated patients with bone metastases andbisphosphonate/denosumab naïve patients with no known bone metastaseswere administered 100 mg Compound 1 daily. NT^(x) plasma levels droppedin the majority of patients relative to baseline based on plasma samplesanalyzed at week 12 of the study. The results indicate the ability ofCompound 1 to inhibit bone resorption.

Study 3: Effect of Compound 1 Administration on ALP Plasma Concentration

The effects of Compound 1 treatment on osteoclast activity was alsoinvestigated based on the measurement of changes in the plasmaconcentration of the bone marker alkaline phosphatase (ALP) in CRPCpatients (N=61). Bisphosphonate/denosumab treated patients with bonemetastases and bisphosphonate/denosumab naïve patients with no knownbone metastases were administered 100 mg Compound 1 daily. ALP plasmalevels dropped in the majority of patients relative to baseline based onplasma samples analyzed at week 12 of the study. The results indicatethe ability of Compound 1 to inhibit bone resorption.

Study 4: Effect of Compound 1 Administration on Circulating Tumor CellPlasma Concentration

The effects of Compound 1 treatment on circulating tumor cell (CTC)concentration in CRPC patients. Patients with CRPC (N=59) wereadministered 100 mg Compound 1 daily. CTC plasma levels dropped in themajority of patients relative to baseline based on samples analyzed atweek 6 or 12 of the study. The results indicate the ability of Compound1 to inhibit tumor cell proliferation.

Study 5: Effect of Compound 1 Administration on CT^(x) PlasmaConcentration

The effects of Compound 1 treatment on osteoclast activity was alsoinvestigated based on the measurement of changes in plasma concentrationof Cross-linked C-terminal telopeptides of type-1 collagen (CT^(x))concentration in bisphosphonate treated and bisphosphonate naïvepatients with CRPC that exhibited bone metastases (N=46). Cr levelsdropped in the majority of patients relative to baseline based on plasmasamples analyzed by ELISA at weeks 6 and 12 of the study. The resultsindicate the ability of Compound 1 to inhibit bone resorption.

Other Embodiments

The foregoing disclosure has been described in some detail by way ofillustration and example, for purposes of clarity and understanding. Theinvention has been described with reference to various specific andpreferred embodiments and techniques. However, it should be understoodthat many variations and modifications can be made while remainingwithin the spirit and scope of the invention. It will be obvious to oneof skill in the art that changes and modifications can be practicedwithin the scope of the appended claims. Therefore, it is to beunderstood that the above description is intended to be illustrative andnot restrictive.

The scope of the invention should, therefore, be determined not withreference to the above description, but should instead be determinedwith reference to the following appended claims, along with the fullscope of equivalents to which such claims are entitled.

1. A method for treating osteolytic bone metastases associated withprostate cancer, comprising administering a compound that duallymodulates MET and VEGF to a patient in need of such treatment, whereinthe compound is the compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein: R¹ is halo; R²is halo; R³ is (C₁-C₆)alkyl; R⁴ is (C₁-C₆)alkyl; and Q is CH or N. 2.The method of claim 1, wherein the prostate cancer is CRPC.
 3. Themethod of claim 2, wherein the dual MET and VEGF modulator is a compoundof Formula Ia

or a pharmaceutically acceptable salt thereof, wherein: R¹ is halo; R²is halo; and Q is CH or N.
 4. The method of claims 1-3, wherein thecompound of Formula I is Compounds 1:

or a pharmaceutically acceptable salt thereof.
 5. The method of claim 4,which isN-(4-{[6,7-bis(methyloxy)quinolin-4-yl]oxy}phenyl)-N′-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide.6. The method of claims 1-5, wherein the compound of Formula (I),Formula I(a) and Compound I is the (L)- or (D)-malate salt.
 7. Themethod of claims 1-6, wherein the compound of Formula (I) is in thecrystalline N-1 form of the (L) malate salt and/or the (D) malate salt.8. The method of claims 1-7 wherein the compound of Formula I, I(a), orCompound 1, or a pharmaceutically acceptable salt thereof, isadministered as a pharmaceutical composition additionally comprising apharmaceutically acceptable carrier, excipient, or diluent.
 9. A methodfor treating osteolytic bone metastases associated with CRPC, comprisingadministering a pharmaceutical composition comprising Compound ofFormula I or the malate salt of Compound of Formula I or anotherpharmaceutically acceptable salt of Compound of Formula I, to a patientin need of such treatment.
 10. A method for reducing bone pain due toosteolytic bone metastases associated with CRPC, comprisingadministering a therapeutically effective amount of a pharmaceuticalcomposition comprising Compound of Formula I or the malate salt ofCompound of Formula I or another pharmaceutically acceptable salt ofCompound of Formula I, to a patient in need of such treatment.
 11. Amethod for treating or minimizing bone pain due to osteolytic bonemetstases associated with CRPC, comprising administering atherapeutically effective amount of a pharmaceutical compositioncomprising Compound of Formula I or the malate salt of Compound ofFormula I or another pharmaceutically acceptable salt of Compound ofFormula I, to a patient in need of such treatment.
 12. A method forpreventing osteolytic bone metastases associated with CRPC, comprisingadministering a therapeutically effective amount of a pharmaceuticalcomposition comprising Compound of Formula I or the malate salt ofCompound of Formula I or another pharmaceutically acceptable salt ofCompound of Formula I, to a patient in need of such treatment.
 13. Amethod for preventing osteolytic bone metastases in patients withprostate cancer who are castration resistant but have not yet advancedto metastatic disease, comprising administering a therapeuticallyeffective amount of a pharmaceutical composition comprising Compound ofFormula I or the malate salt of Compound of Formula I or anotherpharmaceutically acceptable salt of Compound of Formula I, to a patientin need of such treatment.
 14. A method for extending the overallsurvival in patients with CRPC, comprising administering atherapeutically effective amount of a pharmaceutical compositioncomprising Compound of Formula I or the malate salt of Compound ofFormula I or another pharmaceutically acceptable salt of Compound ofFormula I, to a patient in need of such treatment.
 15. A method fortreating osteolytic bone metastases associated with prostate cancer,comprising administering a compound that dually modulates MET and VEGFto a patient in need of such treatment.